中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
1期
63-66
,共4页
黄官梦茜%蒋米尔%黄英%李维敏%陆信武%黄新天%陆民
黃官夢茜%蔣米爾%黃英%李維敏%陸信武%黃新天%陸民
황관몽천%장미이%황영%리유민%륙신무%황신천%륙민
氧化低密度脂蛋白%巨噬细胞%增殖%巨噬细胞迁移抑制因子%核因子-κB通路
氧化低密度脂蛋白%巨噬細胞%增殖%巨噬細胞遷移抑製因子%覈因子-κB通路
양화저밀도지단백%거서세포%증식%거서세포천이억제인자%핵인자-κB통로
Oxidized low density lipoprotein%Macrophage%Proliferation%Macrophage migration inhibitory factor%Nuclear factor-κB pathway
目的 观察氧化低密度脂蛋白(oxLDL)对巨噬细胞增殖和分泌巨噬细胞迁移抑制因子(MMIF/MIF)的作用.方法 (1)制备人单核细胞来源巨噬细胞(MDMs),25、50和75 mg/L浓度oxLDL培养12、24、48 h后,细胞计数试剂盒(CCK-8)法检测细胞增殖,酶联免疫吸附法测定上清MIF浓度;不含oxLDL培养为正常对照(NC)组.(2)制备含稳转核因子-κB (NF-κB)-LUC质粒的MDMs,同法培养,荧光素酶底物(Luciferase)检测细胞NF-κB通路活性.(3)含NF-κB-LUC质粒的MDMs中加入终质量浓度为0、10 μmol/L的NF-κB通路抑制剂BAY 11-7082,同法培养并检测上清MIF浓度.结果 (1)与NC组比较,25、50、75 mg/L oxLDL组12 h时MDMs增殖显著升高,48 h时分别达到17.7％、27.8％和41.2％ (P ＜0.01);其上清液MIF也在12h起升高,48 h时分别是NC组的1.49、1.67和2.09倍(P＜0.01).(2)与NC组比较,oxLDL各组在12h即可检测到NF-κB通路明显激活,24h时MDMs内NF-κB通路活性分别增加38.1％、60.3％和61.1％ (P＜0.01).(3)加入BAY11-7082后,各组上清液MIF浓度在12 h即出现明显下降,48 h时分别下降58.6％、69.3％、69.7％和73.5％ (P＜0.01).结论 25 ～ 75 mg/L浓度的oxLDL可促进MDMs增殖和MIF的分泌,其作用随oxLDL作用时间的延长和浓度的增高而增强.oxLDL可能通过诱导激活NF-κB信号通路促进MDMs合成分泌MIF.
目的 觀察氧化低密度脂蛋白(oxLDL)對巨噬細胞增殖和分泌巨噬細胞遷移抑製因子(MMIF/MIF)的作用.方法 (1)製備人單覈細胞來源巨噬細胞(MDMs),25、50和75 mg/L濃度oxLDL培養12、24、48 h後,細胞計數試劑盒(CCK-8)法檢測細胞增殖,酶聯免疫吸附法測定上清MIF濃度;不含oxLDL培養為正常對照(NC)組.(2)製備含穩轉覈因子-κB (NF-κB)-LUC質粒的MDMs,同法培養,熒光素酶底物(Luciferase)檢測細胞NF-κB通路活性.(3)含NF-κB-LUC質粒的MDMs中加入終質量濃度為0、10 μmol/L的NF-κB通路抑製劑BAY 11-7082,同法培養併檢測上清MIF濃度.結果 (1)與NC組比較,25、50、75 mg/L oxLDL組12 h時MDMs增殖顯著升高,48 h時分彆達到17.7％、27.8％和41.2％ (P ＜0.01);其上清液MIF也在12h起升高,48 h時分彆是NC組的1.49、1.67和2.09倍(P＜0.01).(2)與NC組比較,oxLDL各組在12h即可檢測到NF-κB通路明顯激活,24h時MDMs內NF-κB通路活性分彆增加38.1％、60.3％和61.1％ (P＜0.01).(3)加入BAY11-7082後,各組上清液MIF濃度在12 h即齣現明顯下降,48 h時分彆下降58.6％、69.3％、69.7％和73.5％ (P＜0.01).結論 25 ～ 75 mg/L濃度的oxLDL可促進MDMs增殖和MIF的分泌,其作用隨oxLDL作用時間的延長和濃度的增高而增彊.oxLDL可能通過誘導激活NF-κB信號通路促進MDMs閤成分泌MIF.
목적 관찰양화저밀도지단백(oxLDL)대거서세포증식화분비거서세포천이억제인자(MMIF/MIF)적작용.방법 (1)제비인단핵세포래원거서세포(MDMs),25、50화75 mg/L농도oxLDL배양12、24、48 h후,세포계수시제합(CCK-8)법검측세포증식,매련면역흡부법측정상청MIF농도;불함oxLDL배양위정상대조(NC)조.(2)제비함은전핵인자-κB (NF-κB)-LUC질립적MDMs,동법배양,형광소매저물(Luciferase)검측세포NF-κB통로활성.(3)함NF-κB-LUC질립적MDMs중가입종질량농도위0、10 μmol/L적NF-κB통로억제제BAY 11-7082,동법배양병검측상청MIF농도.결과 (1)여NC조비교,25、50、75 mg/L oxLDL조12 h시MDMs증식현저승고,48 h시분별체도17.7％、27.8％화41.2％ (P ＜0.01);기상청액MIF야재12h기승고,48 h시분별시NC조적1.49、1.67화2.09배(P＜0.01).(2)여NC조비교,oxLDL각조재12h즉가검측도NF-κB통로명현격활,24h시MDMs내NF-κB통로활성분별증가38.1％、60.3％화61.1％ (P＜0.01).(3)가입BAY11-7082후,각조상청액MIF농도재12 h즉출현명현하강,48 h시분별하강58.6％、69.3％、69.7％화73.5％ (P＜0.01).결론 25 ～ 75 mg/L농도적oxLDL가촉진MDMs증식화MIF적분비,기작용수oxLDL작용시간적연장화농도적증고이증강.oxLDL가능통과유도격활NF-κB신호통로촉진MDMs합성분비MIF.
Objective To investigate the effect of oxidized low density lipoprotein (oxLDL) on macrophages and their secretion of macrophage migration inhibitory factor (MMIF/MIF).Methods (1)Preparation of monocytes derived macrophages (MDMs),co-culture with oxLDL at concentrations of 25,50,75 mg/L,respectively.MDMs cultured without oxLDL were normal controls (NC).MDM proliferation was detected using cell counting kit-8 assay (CCK-8 assay) ; supernatant MIF was examined using enzyme linked immunosorbent assay (ELISA).(2) Preparation of MDMs derived from THP-1 cell lines which were transfected with nuclear factor-κB (NF-κB)-LUC plasmids,co-culture with oxLDL as above-mentioned,NF-κB pathway activity was tested by luciferase test.(3) MDMs containing NF-κB-LUC plasmids were co-cultured with oxLDL,self-control study was performed at the same concentration of oxLDL,with or without adding NF-κB pathway inhibitor BAY 11-7082 at the concentration of 10 μmol/L to culture medium.Supernatant MIF was detected.Results (1) Compared with NC,significantly increased MDM proliferation occurred 12 hours after co-culture with oxLDL,the proliferation increased by 17.7％,27.8％ and 41.2％,respectively at 48 hours (P ＜ 0.01) ; supernatant MIF level increased simultaneously,reaching 1.49,1.67 and 2.09 folds,respectively at 48 hours,as compared with NC (P＜0.01).(2)Compared with NC,significant activation of NF-κB pathway was detected 12 hours after co-culture with oxLDL,the activation was increased by 38.1％,60.3％ and 61.1％,respectively at 48 hours (P ＜0.01).(3) Compared with NF-κB pathway inhibitor BAY 11-7082-free group,supematant MIF level decreased significantly at 12 hours in BAY 11-7082 group,it decreased by 58.6％,69.3％,69.7％ and 73.5％,respectively at 48 hours (P ＜ 0.01).Conclusion oxLDL at concentrations of 25 to 75 mg/L could promote MDM proliferation and MIF secretion,this effect was enhanced with prolonged co-culture time.The increased MIF secretion induced by oxLDL might be achieved through activation of the NF-κB pathway.
